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CaMKII regulates diacylglycerol lipase-α and striatal endocannabinoid signaling

Nature Neuroscience volume 16, pages 456–463 (2013)Cite this article

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Abstract

The endocannabinoid 2-arachidonoylglycerol (2-AG) mediates activity-dependent depression of excitatory neurotransmission at central synapses, but the molecular regulation of 2-AG synthesis is not well understood. Here we identify a functional interaction between the 2-AG synthetic enzyme diacylglycerol lipase-α (DGLα) and calcium/calmodulin dependent protein kinase II (CaMKII). Activated CaMKII interacted with the C-terminal domain of DGLα, phosphorylated two serine residues and inhibited DGLα activity. Consistent with an inhibitory role for CaMKII in 2-AG synthesis, in vivo genetic inhibition of CaMKII increased striatal DGL activity and basal levels of 2-AG, and CaMKII inhibition augmented short-term retrograde endocannabinoid signaling at striatal glutamatergic synapses. Lastly, blockade of 2-AG breakdown using concentrations of JZL-184 that have no effect in wild-type mice produced a hypolocomotor response in mice with reduced CaMKII activity. These findings provide mechanistic insights into the molecular regulation of striatal endocannabinoid signaling with implications for physiological control of motor function.

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Figure 1: DGLα interacts with CaMKII.
Figure 2: CaMKIIα selectively phosphorylates DGLα at Ser782 and Ser808.
Figure 3: CaMKII inhibits DGL activity.
Figure 4: Inhibiting CaMKII or preventing CaMKII autophosphorylation at Thr286 enhances endocannabinoid-mediated retrograde transmission in striatal MSNs.
Figure 5: Inhibition of 2-AG breakdown selectively suppresses motor activity in Camk2aT286A knock-in mice (KI).

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Acknowledgements

This work was supported by US National Institutes of Health (T32-NS007491 and T32-MH065215 to B.C.S.; K01-NS073700 to A.J.B.; K05-DA021696 and R01-DA011322 to K.M.; R01-AA019455 to D.G.W.; K08-MH090412 to S.P.; and R01-MH063232 and R01-NS078291 to R.J.C.), the Michael J. Fox Foundation (R.J.C.) and the Luton Society (S.P.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the US National Institutes of Health. Behavioral studies were performed in the Murine Neurobehavior Core and mass spectrometry and proteomics studies were performed at the Mass Spectrometry Research Center, both at Vanderbilt University Medical Center. We thank B. Cravatt (The Scripps Research Institute) for plasmids encoding DGLα-V5 and DGLβ-V5.

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Authors and Affiliations

  1. Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Brian C Shonesy, Tyler Rentz, Anthony J Baucum II, Nidhi Jalan-Sakrikar, Danny G Winder, Sachin Patel & Roger J Colbran

  2. Vanderbilt Brain Institute, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Brian C Shonesy, Xiaohan Wang, Teniel S Ramikie, Victoria S Cavener, Danny G Winder, Sachin Patel & Roger J Colbran

  3. Vanderbilt International Scholar Program, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Xiaohan Wang

  4. Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Kristie L Rose

  5. Department of Psychiatry, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Teniel S Ramikie & Sachin Patel

  6. The Gill Center and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana, USA

    Ken Mackie

  7. Vanderbilt–Kennedy Center for Research on Human Development, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Danny G Winder, Sachin Patel & Roger J Colbran

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Contributions

B.C.S. and A.J.B. performed all of the immunoprecipitation studies and LC/MS quantification of striatal lipid levels. B.C.S. performed all of the DGL activity assays. V.S.C. carried out the GST cosedimentation assay. B.C.S. and X.W. generated the GST-fusion proteins and DGLα mutants, and performed the phosphorylation site identification studies. B.C.S. and K.L.R. performed the LC/MS analysis of phosphorylation sites. B.C.S., T.S.R. and S.P. conducted the electrophysiology experiments. B.C.S. and T.R. carried out the behavioral experiments. B.C.S. analyzed data from all experiments. N.J.-S. performed the T286 phosphorylation measurements in HEK293 cells. K.M. contributed the DGL antibodies. B.C.S., D.G.W., S.P. and R.J.C. designed the experiments. B.C.S., S.P. and R.J.C. prepared the manuscript with input from all other authors.

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Correspondence to Sachin Patel or Roger J Colbran.

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Shonesy, B., Wang, X., Rose, K. et al. CaMKII regulates diacylglycerol lipase-α and striatal endocannabinoid signaling. Nat Neurosci 16, 456–463 (2013). https://doi.org/10.1038/nn.3353

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